Heparinase, isolated from Flavobacterium heparinum, has been shown to be effective in neutralizing the anticoagulant effect of heparin in plasma samples in vitro. It has also been shown to shorten the activated partial thromboplastin time of heparinized rabbits when injected in vivo. We propose in this project to study the various parameters of growth of F. heparinum which might affect the yield of organisms or the yield of heparinase. Our object will be to maximize the yield of heparin-degrading enzymes, and minimize the yield of contaminants which have been shown in crude extracts to prolong the activated partial thromboplastin time. Once production of enzymes on a large scale and a regular basis is assured, we propose to demonstrate in the Clinical Coagulation Laboratory at North Carolina Memorial Hospital, that heparinase is useful in determining whether a prolonged partial thromboplastin time or prothrombin time is due to the presence of heparin, or due rather to another coagulation defect requiring further definition. We wish further to determine if heparinase-treated plasma samples will permit quantitative determination of individual coagulation factors. Finally, those coagulation tests validated in our laboratories, especially the use of heparinase as an initial screening test for heparin contamination of plasma samples, will be demonstrated in the laboratories affiliated with the University of North Carolina through the Area Health Education Center program.